Enhancement of the mechanical properties of HDPE mineral nanocomposites by filler particles modulation of the matrix plastic/elastic behavior
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Nanotechnology Reviews 2022; 11: 312–320
Research Article
Yousef Murtaja, Lubomír Lapčík*, Harun Sepetcioglu, Jakub Vlček, Barbora Lapčíková,
Martin Ovsík, and Michal Staněk
Enhancement of the mechanical properties of
HDPE mineral nanocomposites by filler particles
modulation of the matrix plastic/elastic behavior
https://doi.org/10.1515/ntrev-2022-0023
received September 20, 2021; accepted November 26, 2021
1 Introduction
Abstract: Two different nanosized mineral fillers (nano The study of the effect of the nano/microsized mineral
calcium carbonate and nanoclay) were used in the high fillers blended in the high density poly(ethylene) (HDPE)
density poly(ethylene) (HDPE) composites pilot plant composite matrix on the mechanical properties of the pre-
production. Structural and mechanical properties of the pared composites gained excessive attraction in the past
prepared composites were examined in this study. The 10 years due to their wide application in automotive, aero-
homogenous filler distribution was confirmed in the tested space industries [1,2], formulation engineering [3], sound
samples by scanning electron microscopy, transmission damping materials [4], highly conductive polymeric nano-
electron microscopy, and energy dispersive spectroscopy composites [5], dielectric material applications [6], etc.
analyses. The fillers’ fortifying effect on polymer compos- Semi-crystalline polymers exhibit, in general, a free-phase
ites’ mechanical performance was confirmed as indicated continuum system with the crystalline and amorphous
by the increased elastic modulus and indentation modulus. phases separated with interphase. The crystalline part is
Additionally, the possible modulation of the plastic-elastic formed with mutually connected spherulites consisting of
mechanical behavior was confirmed by the type of the filler crystalline lamellae dispersed in the amorphous phase [7].
as well as its concentration used in the final composites The type, shape, and size of the mineral filler have a strong
testing articles. impact on the mechanical properties of the thermoplastic-
Keywords: nanosized mineral fillers, HDPE, composites, polymer-based composites as well as on their melting
mechanical properties behavior and crystallization kinetics [8]. Furthermore,
the nature and the quality of the mutual adhesion between
the filler and the polymer matrix [9], filler particle size,
shape, and particle size distribution have a paramount
* Corresponding author: Lubomír Lapčík, Department of Physical
Chemistry, Faculty of Science, Palacky University, 17. Listopadu 12, effect on the final composite application performance
771 46 Olomouc, Czech Republic; Department of Food Technology, [10,11]. This study offers the mechanical testing of the com-
Faculty of Technology, Tomas Bata University in Zlin, Nam. T.G. posites prepared from the commercial fillers compounded
Masaryka 275, 760 01 Zlin, Czech Republic, with HDPE in industrial-scale semi-pilot conditions. The
e-mail: lapcikl@seznam.cz
main aim was to confirm the large-scale processability
Yousef Murtaja, Jakub Vlček: Department of Physical Chemistry,
Faculty of Science, Palacky University, 17. Listopadu 12, 771 46
and reproducibility of the manufacturing steps during com-
Olomouc, Czech Republic posites production.
Harun Sepetcioglu: Department of Metallurgy and Materials
Engineering, Selçuk University, Faculty of Technology, Konya 42075,
Turkey
Barbora Lapčíková: Department of Physical Chemistry, Faculty of 2 Materials
Science, Palacky University, 17. Listopadu 12, 771 46 Olomouc,
Czech Republic; Department of Food Technology, Faculty of HDPE of HD8100M grade used in our entire experiment
Technology, Tomas Bata University in Zlin, Nam. T.G. Masaryka 275,
was supplied by Polymer Marketing Company Limited
760 01 Zlin, Czech Republic
Martin Ovsík, Michal Staněk: Department of Manufacturing
(Thailand). The density of the resin was 0.952 g/cm3
Technology, Faculty of Technology, Tomas Bata University in Zlin, with a melt flow index of 0.25 g/10 min. The nano calcium
Nam. T.G. Masaryka 275, 760 01 Zlin, Czech Republic carbonate (CaCO3) particles, also known as adaCAL-N1-C,
Open Access. © 2022 Yousef Murtaja et al., published by De Gruyter. This work is licensed under the Creative Commons Attribution 4.0
International License.
Modulation of the mechanical properties of HDPE mineral nanocomposites 313 Figure 1: TEM images of the fillers used in this study: Nanoclay and nano CaCO3.
314 Yousef Murtaja et al.
were received from Adacal Co. (Turkey) and were treated pure diiodomethane of ACS reagent grade (Sigma Aldrich,
with stearic acid prior to further processing. Particles’ average Germany) were used as wetting liquids for contact angle
size d50 was of 0.05 µm as obtained from scanning electron measurements.
microscopy (SEM) measurements. Nanoclay particles (i.e.,
EsanNANO 1-140) were supplied by EczacıbasıEsan (Turkey).
Particles’ average size d50 was of 2.7 µm as obtained from laser
diffractometer measurements. SEM and transmission electron
3 Methods
microscopy (TEM) images of the prepared samples showed
that the nanofillers were homogeneously dispersed within 3.1 SEM and TEM
the HDPE matrix (Figure 1). This fact was confirmed also by
the energy dispersive spectroscopy (EDS) mapping study. TEM (FEI Tecnai G2 Spirit Biotwin model, FEI Company,
The chemical compositions and physical properties of USA) was used to characterize the shape and morphology
the used nano calcium carbonate as well as that of the nano- of the filler particles. TEM images were taken by placing
clay are given in Table 1 and are also available in refs [12,13]. nanofiller samples on a standard 400 grid copper mesh.
Images of both fillers captured by TEM are shown in Dispersions of acetone fillers were ultrasonicated for
Figure 1. For testing, the set of samples of different filler 15 min and were casted on the copper mesh and air dried.
concentrations were prepared and labelled as CC for nano TEM measurements were performed at 120 kV acceler-
calcium carbonate and NC for nanoclay. ating voltage. Nano CaCO3 and nanoclay’s distributions
Nanoclay/HDPE nanocomposites were prepared by in HDPE matrix were analyzed by TEM. An ultra-thin
melt mixing system composed of Banbury mixer, single section of about 100 nm thickness were cut from filled
screw extruder, and granule cutting unit allowing semi- samples using a microtome device (CM1950) supplied
pilot production in 100 kg scale. The processing tempera- by Leica Microsystems Inc. (Buffalo Grove, USA) in a
ture in the mixer was kept at 180°C and the temperature low-temperature environment. For further examination
was reached in 15 min after filling the mixer chamber with of the distribution of nanofillers, the composites were
both the HDPE granules and the filler [14]. The apparatus characterized by SEM using a Zeiss EvoLS10 equipped
was then followed by the extruder with a conveyor belt with an energy-dispersive X-ray detector (Germany).
and the cutting unit. Single screw extruder was operating
at 330 rpm screw speed with five barrel temperature pro-
files of 200, 190, 190, 190, and 220°C. Nanoclay/HDPE 3.2 Thermal analysis
nanocomposite hot mixtures were cut in water into the
shape of granules. Then, they were molded as tensile and Differential scanning calorimetry (DSC) experiments were
impact test specimens using a PS40E5ASE injection mold- performed according to ASTM E1356 by using a TA Instrument
ing machine with a melt temperature of 210°C, mold tem- S10 model (Waters, USA) at a nitrogen flow rate of 50 mL/min.
perature of 65°C, and injection pressure of 50 MPa [14]. Virgin HDPE and its nanocomposites’ glass transition tem-
Similarly, the nano CaCO3/HDPE nanocomposites were pre- peratures (Tg) were determined from DSC curves by means
pared by the melt mixing method as well by use of the same of the midpoint method at 10°C/min heating rate from 30 to
compounder system and the processing parameters as men- 300°C [13,15].
tioned above in the case of nanoclay/HDPE composites
preparation. For both fillers, the weight ratios of the HDPE
and the fillers were maintained to obtain the samples of 3.3 Uniaxial tensile testing
the given filler weight concentration such as 1, 3, 5, 10,
and 15 wt% for CC/HDPE composites and 1, 2, 3, 4, and Universal Testing Machine Autograph AGS-100 Shimadzu
5 wt% for NC/HDPE composites. (Japan) and Zwick 1456 multipurpose tester (Germany)
Millipore water (USA) with a conductivity of 0.06 equipped with Compact Thermostatic Chamber TCE Series
µS/cm, ethylene glycol p.a., (Lach-Ner, Czechia), and 99% were used for tensile testing of injection-molded specimens.
Table 1: Physicochemical properties of applied fillers: nanoclay and nano CaCO3
Filler type Color Density (g/cm3) Surface area (g/m2) Particle size (μm)
Nanoclay Ultra white 1.98 19 2–20
Nano CaCO3 Ultra white 2.95 28 0.05–0.10Modulation of the mechanical properties of HDPE mineral nanocomposites 315
Figure 2: (a) General mechanical behavior of NC/HDPE and (b) CC/HDPE composites with different filler concentrations from tensile testing
experiments as obtained for 50 mm/min deformation rates at ambient temperature expressed as stress vs strain dependencies.
All data ptwere recorded as per CSN EN ISO 527-1 and CSN EN 179-2 standard, allowing 25 J energy drop. Each experi-
ISO 527-2 standards taking the tested gauge length of 8 cm. All ment was repeated 10×, and the mean values and stan-
experiments were performed at room temperature up to break dard deviations were calculated. All experiments were
with 50, 100, and 200 mm/min deformation rates. Strength at performed at the laboratory ambient conditions of 25°C
break, Young’s modulus, and strain at break were obtained temperature.
from the stress-strain dependence plot(s). Each experiment
was repeated 10×, and the mean values and standard devia-
tions were calculated. All experiments were performed at the
3.5 Surface free energy (SFE)
laboratory ambient conditions of 25°C temperature.
characterization
3.4 Charpy impact testing The SFE of the studied composites and pure HDPE was
determined by the static contact angle of wetting mea-
Impact tests were carried out using Zwick 513 Pendulum surements based on axisymmetric drop shape analysis.
Impact Tester (Germany) according to the CSN EN ISO All measurements were performed at 23°C and repeated
Figure 3: Young’s modulus and filler concentration dependencies of the CC/HDPE and NC/HDPE composites obtained with tensile testing
experiments for different deformation rates.316 Yousef Murtaja et al.
Table 2: Results of the tensile testing experiments of the studied HDPE composites at 50, 100, and 200 mm/min deformation rates. Filler concentrations indicated in the sample description are
7× with a Krüss DSA 30 (Krüss, Germany). The Owens,
Fracture toughness (kJ/m2)
Wendt, Rabel, and Kaelble extended Fowkes theory was
used to calculate the SFE of the tested composites and
pure HDPE from the average static contact angles for
28.44 ± 0.09
water, ethylene glycol, and diiodomethane [16,17].
26.29 ± 1.04
22.06 ± 1.42
31.48 ± 2.43
17.26 ± 0.92
23.35 ± 0.27
31.79 ± 3.94
36.69 ± 5.15
29.21 ± 2.21
28.69 ± 3.4
24.37 ± 1.0
3.6 Micro hardness
26.6 ± 2.2
16.6 ± 0.6
16.4 ± 6.0
19.8 ± 0.3
14.4 ± 5.9
20.5 ± 2.1
14.1 ± 0.4
22.9 ± 1.5
12.1 ± 0.6
11.2 ± 2.2
17.2 ± 3.1
Micro-indentation tests were performed on a micro-inden-
200
tation tester (Micro Combi Tester, Anton Paar, Austria),
Strain at break (%)
according to the CSN EN ISO 14577 standard. The applied
Rate (mm/min)
28.5 ± 14.7
20.5 ± 10.5
32.88 ± 10
31.1 ± 11.2
33.2 ± 6.7
25.8 ± 5.0
20.5 ± 7.4
diamond tip was of the cube corner shape (Vickers, Anton
19.3 ± 4.6
19.7 ± 6.9
35.7 ± 7.2
19.8 ± 1.0
Paar, Austria). Measurement parameters were set as fol-
100
lows: maximum load of 3 N, loading rate (unloading rate)
of 6 N/min, and holding time of 90 s. All experiments were
32.8 ± 15.9
22.0 ± 0.8
performed according to the depth-sensing indentation
28.9 ± 2.7
25.0 ± 3.0
27.6 ± 2.2
20.1 ± 2.6
30.8 ± 3.1
18.6 ± 2.3
19.8 ± 3.2
20.5 ± 0.1
21.2 ± 8.7
method, allowing simultaneous measurement of the
acting force on the indentor and the displacement of the
50
indentor’s tip. The indentation hardness (HIT) was calcu-
lated as the maximum load (Fmax) on the projected area of
30.0 ± 2.8
22.4 ± 0.4
24.7 ± 0.8
25.6 ± 0.4
26.3 ± 0.7
24.7 ± 0.2
27.4 ± 2.4
28.1 ± 2.8
27.9 ± 2.5
25.1 ± 0.5
25.0 ± 1.1
the hardness impression (Ap). Indentation modulus (EIT)
200
was calculated from the plane strain modulus of elasticity
(E*) using an estimated Poisson’s ratio (ν) of the samples
Upper yield (MPa)
Rate (mm/min)
(0.3–0.4 [18,19]):
23.9 ± 0.3
25.0 ± 0.2
27.4 ± 2.2
25.4 ± 3.4
23.5 ± 0.2
26.5 ± 0.5
23.9 ± 1.6
25.2 ± 1.0
24.8 ± 1.1
24.3 ± 1.1
25.3 ± 1.1
Fmax
HIT = , (1)
100
Ap
EIT = E ⁎(1 − ν 2 ) . (2)
24.4 ± 0.4
24.6 ± 0.2
25.4 ± 0.6
23.6 ± 0.5
25.8 ± 3.2
23.3 ± 0.1
24.3 ± 1.3
27.3 ± 1.9
23.6 ± 1.5
24.5 ± 1.1
23 ± 0.2
Each measurement was repeated 10×, and mean values
50
and standard deviations were calculated. All experiments
were performed at the laboratory ambient conditions of
1,328.3 ± 78.0
1,099.2 ± 16.2
1,215.6 ± 48.8
1,223.3 ± 25.0
1,202.7 ± 32.1
1,150.5 ± 23.0
1,134.9 ± 61.2
1,237.9 ± 13.7
1,167.7 ± 23.7
1,316.4 ± 0.4
25°C temperature.
1,360.7 ± 1.7
200
4 Results and discussion
Young’s modulus (MPa)
Rate (mm/min)
1,,034.3 ± 299
1,070.9 ± 303
943.8 ± 142.2
1,018.8 ± 246
1,060.1 ± 240
1,112.8 ± 273
648.2 ± 419
982.9 ± 217
982.7 ± 213
Results of the tensile testing experiments of the studied
1,368 ± 2.5
1155 ± 255
composites are shown in Figure 2. Obtained stress vs strain
deformation dependencies exhibited typical patterns corre-
100
sponding to the elastic region (does not exceed 3% strain for
small deformations), elastic-plastic transition region (does
1,304.9 ± 83.7
1,288.9 ± 54.7
1,248.7 ± 28.1
1,121.4 ± 26.8
1,265.4 ± 51.9
1,465.1 ± 12.3
970.1 ± 111.7
837.5 ± 10.8
1,470 ± 54.7
1,322 ± 10.8
1,255 ± 11.9
not exceed 10% strain for small deformations), and the stress
plateau draw region occurred for 5 wt% nanoclay/HDPE
composites and 15 wt% for CC/HDPE composites at strains
50
exceeding 18% (NC/HDPE composites) and 12% (CC/HDPE
given in wt%
composites), respectively [20]. Similar dependencies were
CC_10%
CC_15%
Sample
NC_4%
NC_2%
NC_3%
NC_5%
CC_3%
CC_5%
NC_1%
CC_1%
found in our previous studies for industrial HDPE mineral
HDPE
composites at the same deformation rate [13,15]. Pure HDPEModulation of the mechanical properties of HDPE mineral nanocomposites 317 Figure 4: Upper yield and filler concentration dependencies of the CC/HDPE and NC/HDPE composites obtained with tensile testing experiments for different deformation rates. Figure 5: Strain at break and filler concentration dependencies of the CC/HDPE and NC/HDPE composites obtained with tensile testing experiments for different deformation rates. exhibited more stiff tensile deformation behavior in the absence for both mineral fillers, as shown in Figure 3. Such behavior of the stress plateau draw region. In contrary to that CC/HDPE was confirmed for all applied deformation rates, as given composites, were characteristic of more elasto-plastic beha- in Table 2. For example, the absolute value of the modulus vior as reflected for 1 wt% filler concentration (Figure 1b). of elasticity of 970.1 ± 111.7 MPa for original HDPE was In the case of NC/HDPE composites, there is a significant increased to 1470.0 ± 54.7 MPa by about 51.5% in the influence of elastic-plastic properties. With increasing case of CC/HDPE composites with 5 wt% filler con- filler concentration, greater plastic behavior was observed, centration. For the NC/HDPE composites, the modulus as reflected by the increased stress plateau draw region, as E was increased by about 34.5% to the absolute value shown in Figure 2b. The abovementioned behavior was also of 1304.9 ± 83.7 MPa with 4 wt% filler concentration. These accompanied by the corresponding increase in the Young’s corresponded very well with previously published data that modulus of elasticity with increasing filler concentration stiff mineral filler particles were responsible for the observed
318 Yousef Murtaja et al.
Figure 7: Indentation modulus (EIT) vs filler concentration depen-
dencies. Filler type: full circle – Nano CaCO3 and empty
diamond – Nanoclay. *Point omitted for linear regression.
MPa were increasing up to 26.3 ± 0.7 MPa and up to
Figure 6: Fracture toughness and filler concentration dependencies 28.1 ± 2.8 MPa, respectively.
of the CC/HDPE and NC/HDPE composites as obtained with Charpy The results of the filler concentration strain at break
impact testing. dependencies measured at different deformation rates are
shown in Figure 5. In the case of CC/HDPE composites,
observed trend was such that the strain at break was
Table 3: Total SFEs and their components (polar and dispersive) of
decreased with increasing filler concentration, thus reflecting
the studied HDPE composites calculated by Owens, Wendt, Rabel,
and Kaelble approach from contact angle measurements performed the loss of plastic behavior, indicating more brittle-like
by means of the axisymmetric drop shape analysis (measured at mechanical behavior. This fact fits very well with the well-
23°C temperature) known theory that polymers with higher crystallinity exhibit
higher elastic properties than amorphous systems, which
Sample SFE (mJ/m2) exhibit more plastic behavior [22]. However, the opposite
Total Polar Dispersive trend was observed for the NC/HDPE composites, where
the increased strain at break with increasing filler concentra-
HDPE 19.48 ± 2.91 15.29 ± 2.10 4.19 ± 0.81
CC_1% 36.51 ± 33.80 15.61 ± 14.82 15.61 ± 14.82
tion was found. This was most probably due to the pre-
CC_3% 36.49 ± 14.86 11.84 ± 7.77 24.65 ± 7.08 ferential orientation of the individual platelet shape like
CC_5% 39.70 ± 1.09 0.19 ± 0.46 39.51 ± 0.63 nanoclay filler particles in the polymer macromolecular
CC_10% 23.45 ± 4.41 0.01 ± 0.07 23.44 ± 4.34 chains interphase induced during sample injection molding
CC_15% 26.19 ± 0.05 0.41 ± 0.01 25.78 ± 0.03 processing [20]. The comparison of the obtained magnitudes
NC_1% 30.44 ± 1.04 0.12 ± 0.04 30.32 ± 1.00
of the elongation at break parameter for 5 wt% nanocompo-
NC_2% 30.28 ± 0.92 1.19 ± 0.15 29.09 ± 0.76
NC_3% 32.95 ± 2.58 00.08 ± 0.16 32.86 ± 2.42 site filler concentrations (both the nano-clay as well as of
NC_4% 29.22 ± 0.66 0.22 ± 0.13 29.00 ± 0.53 the nano-calcium carbonate) confirmed higher plasticity
NC_5% 23.45 ± 4.41 0.01 ± 0.07 23.44 ± 4.34 of the composite matrix composed from the nanoclay flat
like filler particles for all applied deformation rates. This
finding was also in agreement with the mutual comparison
steady increase in the modulus of elasticity of polymer- of the observed magnitudes of the Young’s modulus of
based composites with increasing filler content [21,22]. elasticity, where the CC/HDPE samples exhibited higher
Upper yield vs filler concentration dependencies of modulus of elasticity than the NC/HDPE samples.
the HDPE composites are shown in Figure 4. They are Observed fracture toughness vs filler concentration
typical non-linear patterns for both fillers under study dependencies of the tested composites are shown in
(nanoclay and nano calcium carbonate). However, in Figure 6. A decrease in the fracture toughness was found
the case of CC/HDPE composites, the significant changes for all the composites studied in comparison with the
in the upper yield with increasing filler concentration virgin polymer (HDPE). The most significant decrease of
were not observed with the exception of the composites about 52.96% was found for the composites containing
with 4 and 5 wt% filler concentration for the 200 mm/min nano CaCO3 of 3 wt% filler concentration. For NC/HDPE
deformation rate, where the original values of 25.6 ± 0.4 composites, the observed fracture toughness was decreasedModulation of the mechanical properties of HDPE mineral nanocomposites 319
to 22.06 ± 1.42 kJ/m2 magnitude, which was about 39.87% case of nano CC/HDPE composites with 5 wt% filler con-
decrease compared to the virgin HDPE. These results clearly centration. For the NC/HDPE composites, the modulus of
demonstrate the higher brittle character of the composites elasticity was increased by about 34.5% to the absolute
in comparison to the virgin HDPE. value of 1304.9 ± 83.7 MPa with 4 wt% of filler concentra-
It was found by DSC thermal analysis that the HDPE tion. Additionally, there was confirmed possible modula-
filled with nano CaCO3 exhibited higher thermal stability tion of the plastic-elastic mechanical behavior by the type
in comparison with the virgin HDPE as reflected by increasing of the filler as well as its concentration used in the final
Tg from 103.9°C (virgin HDPE) to 126.6°C for 15 wt% CC/HDPE composites testing articles, as confirmed by the increased
composites. These results were in excellent agreement with stress plateau draw region that occurred during the tensile
the published data of Viljoen and Labuschagné [23]. Similarly, testing and decreased elongation at break with increasing
the Tg increased with increasing nano-clay content in the filler concentration. From the practical point of view, the
nano-clay/HDPE composites to 128.2°C, thus confirming that 5 wt% filler concentration seems to be the most favorable
the fillers enhanced interaction with the base HDPE matrix. one for both CC/HDPE as well as nano-clay/HDPE compos-
Results of the SFE calculations are given in Table 3. ites. Simultaneously, the higher thermal stability was
Obtained results indicated that both fillers (CaCO3 as well found for both nanocomposites in comparison to the virgin
as nanoclay) contributed to the observed decrease in the HDPE, thus confirming that the fillers enhanced interac-
originally more polar character of the virgin HDPE to less tion with the HDPE matrix. Based on the abovementioned
polar one as indicated by the observed increase in the conclusions, it seems to be advantageous for the applica-
dispersive part of SFE from 4.19 ± 0.81 mJ/m2 (virgin tion of the latter nanocomposites as the structural ele-
HDPE) to 39.51 ± 0.63 mJ/m2 (5 wt% CC/HDPE compos- ments in the complex product designs offering combined
ites) and to 32.86 ± 2.42 mJ/m2 (3 wt% NC/HDPE compos- elasto-plastic mechanical behavior in the relatively wide
ites). The magnitude of the total SFE was increased from deformation rates regimes accompanied with the higher
19.48 ± 2.91 mJ/m2 (virgin HDPE) by about 103.8% up to thermal stability.
39.70 ± 1.09 mJ/m2 (5 wt% CC/HDPE composites) and by
about 69.18% up to 32.95 ± 2.58 mJ/m2 (3 wt% NC/HDPE Funding information: Financial support from the internal
composite). Observed results indicate further improve- grants of Palacky University in Olomouc (project number
ment in the composites interface’s adhesive properties, IGA_PrF_2021_031) and of Tomas Bata University in Zlin
for example, suitable for coating or adhesive joints tech- (project numbers IGA/FT/2021/004 and IGA/FT/2021/005)
nical applications. are gratefully acknowledged. Financial support to the
The composites micro-indentation testing results are author YM by Fischer scholarship of the Faculty of
shown in Figure 7 where indentation modulus EIT vs filler Science, Palacky University in Olomouc in 2021 year is
concentration patterns are shown. These were character- gratefully acknowledged as well.
istic with the gradual increase in the indentation modulus
with increasing concentrations of both fillers, confirming Author contributions: All authors have accepted respon-
the fortification effect of the fillers on the mechanical prop- sibility for the entire content of this manuscript and
erties of the tested samples. approved its submission.
Conflict of interest: The authors state no conflict of
interest.
5 Conclusion
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